Sp3, single-pot, solid-phase-enhanced sample-preparation樣本處理方法經典文獻參考
2025-5-10 12:35:25點擊：
SP3（single-pot, solid-phase-enhanced sample-preparation）是一種基于順磁顆粒的蛋白質組預處理工作流程，通過有機溶劑乙醇或乙腈驅動驅動蛋白在羧基或其它親水性磁珠表面被捕獲（如圖所示）。標準 SP3流程中，蛋白質通過親水相互作用機制與磁珠結合，結合了蛋白的磁珠被束縛在溶劑化層，可沖洗去除不需要的污染物，然后在一定條件下洗脫純化的蛋白質。這種親水捕獲機制類似于親水相互作用色譜（HILIC）中所涉及的機制，已被廣泛證明是分離多種生物分子的有效工具。廈門普睿邁格生物科技有限公司開發的SP3-Carboxyl磁珠http://www.kismrs.cn/Product/1736805216.html可以完美替代SP3方案中最常使用的是Cytiva的Sera Mag SpeedBeads（GE Healthcare），提供了測定性能與成本之間最佳平衡。
以下是Sp3磁珠在蛋白組學樣本前處理方面的經典文獻，供參考：
- Hughes C S, Foehr S, Garfield D A, et al. Ultrasensitive proteome analysis using paramagnetic bead technology[J]. Molecular systems biology, 2014, 10(10): 757.
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Sielaff M, Kuharev J, Bohn T, et al. Evaluation of FASP, SP3, and iST protocols for proteomic sample preparation in the low microgram range[J]. Journal of proteome research, 2017, 16(11): 4060-4072.
- Moggridge S, Sorensen P H, Morin G B, et al. Extending the compatibility of the SP3 paramagnetic bead processing approach for proteomics[J]. Journal of proteome research, 2018, 17(4): 1730-1740.
- Hughes C S, Moggridge S, Müller T, et al. Single-pot, solid-phase-enhanced sample preparation for proteomics experiments[J]. Nature protocols, 2019, 14(1): 68-85.
- Hughes C S, Sorensen P H, Morin G B. A standardized and reproducible proteomics protocol for bottom-up quantitative analysis of protein samples using SP3 and mass spectrometry[J]. Proteomics for Biomarker Discovery: Methods and Protocols, 2019: 65-87.
- Müller T, Kalxdorf M, Longuespée R, et al. Automated sample preparation with SP 3 for low‐input clinical proteomics[J]. Molecular systems biology, 2020, 16(1): e9111.
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Mikulá?ek K, Kone?ná H, Potě?il D, et al. SP3 protocol for proteomic plant sample preparation prior LC-MS/MS[J]. Frontiers in plant science, 2021, 12: 635550.
- Duong V A, Lee H. Bottom-up proteomics: advancements in sample preparation[J]. International Journal of Molecular Sciences, 2023, 24(6): 5350.
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